Use of 4-Substituted Tetrahvdropyridines for the Manufacture of Medicaments Acting Upon TGF-beta 1
The present invention relates to the use of certain 1,2,3,6-tetrahydropyridine derivatives, and to their pharmaceutically acceptable salts and solvates for the preparation of medicaments capable of increasing the levels of TGF-xcex21 (Transforming growth factor-xcex21).
TGF-xcex21 is a multifunctional and ubiquitous peptide which is constituted, in its active form, by two identical sub-units linked by a disulphide bridge. As illustrated by P. Bedesse and V. Paradis (Journal of Hepatology, 1995, 22, 37-42), TGF-xcex21 has been identified as a factor which induces cell growth in transformed fibroblasts, but many other cell functions have been discovered successively.
The WO 93/09808 application describes the use of TGF-xcex21 for the treatment of damages to the central nervous system.
The WO 96/34881 and WO 94/17099 applications claim novel peptides which have a similar activity to that of TGF-xcex21 and which may be used for the treatment of several pathologies.
TGF-xcex21 is for example implicated in the control of the cell cycle, in angiogenesis, in cellular differentiation, in embryogenesis, in tissue repair, as well as in apoptosis.
Amongst these activities, the anti-apoptotic effect of TGF-xcex21 is very important due to its pharmacological implications.
xe2x80x9cApoptosisxe2x80x9d, or xe2x80x9cprogrammed cell deathxe2x80x9d, indicates the whole of the physiological processes linked to cell death. In its terminal phase, apoptosis is characterised by an activation of the endonucleases which cleave double-stranded DNA in the internucleosomal regions, thus generating mono- and oligo-nucleosomes which complex with histones. An enrichment in oligo- and mono-nucleosomes linked to histones is thus observed in the cytoplasm of the apoptotic cells.
Although this phenomenon is physiological, in contrast to necrosis, it may also be caused by pathological stimulations.
D. A. Carson and J. M. Ribeiro report (The Lancet 1993, 341, 1251-1254) the role of apoptosis in certain pathologies such as immuno-depression, immune deficiencies in patients suffering from AIDS, cell aging, and degenerative illnesses.
J. Mathieu et al. (Ann. pharmaceutiques francaises 1996, 54, 5, 193-201) demonstrated that the pathological effects caused by chemical and physical agents such as free radicals and ion3 sing radiation are caused by the pro-apoptotic effects of these agents.
The apoptosis-regulating products were described in the WO 96/21449 patent application. The general formula includes both inhibitors and stimulators of apoptosis, without the means of distinguishing them from one another being given.
It has now been found that certain tetrahydropyrdines increase circulating and cellular and extracellular levels of TGF-xcex21.
Thus, the object of the present invention is the use of a 4-substituted 1,2,3,6-tetrahydropyridine of formula (I): 
in which:
R1 represents a halogen or a CF3, (C1-C4)alkyl or (C1-C4)alkoxy group;
Y represents a nitrogen atom or a CH group;
Zxe2x80x2 and Zxe2x80x3 each represent hydrogen or a (C1-C3)alkyl group, or one represents hydrogen and the other a hydroxy group, or both, together, represent an oxo group;
Z represents
a phenyl radical;
a phenyl radical monosubstituted with a substituent X, X being
a) a (C1-C6)alkyl; (C1-C6)alkoxy; (C3-C7)carboxyalkyl; (C1-C4)alkoxycarbonyl(C1-C6)alkyl; (C3-C7)carboxyalkoxy or (C1-C4)-alkoxycarbonyl(C1-C6)alkoxy group;
b) a group selected from a (C3-C7)cycloalkyl, (C3-C7)cycloalkyloxy, (C3-C7)cycloalkylmethyl, (C3-C7)cycloalkylamino and cyclohexenyl group, it being possible for said group to be substituted with a halogen, hydroxy, (C1-C4)alkoxy, carboxy, (C1-C4)alkoxycarbonyl, amino, mono- or di-(C1-C4)alkylamino;
c) a group selected from a phenyl, phenoxy, phenylamino, N-(C1-C3)alkylphenylamino, phenylmethyl, phenylethyl, phenylcarbonyl, phenylthio, phenylsulphonyl, phenylsulphinyl or styryl, it being possible for said group to be mono- or poly-substituted on the phenyl group with a halogen, CF3, (C1-C4)alkyl, (C1-C4)alkoxy, cyano, amino, mono- or di-(C1-C4)alkylamino, (C1-C4)acylamino, carboxy, (C1-C4)alkoxycarbonyl, aminocarbonyl, mono- or di-(C1-C4)alkylaminocarbonyl, amino(C1-C4)alkyl, hydroxy(C1-C4)alkyl or halo(C1-C4)alkyl;
a phenyl radical disubstituted with a substituent R2, R2 being a halogen or a hydroxy, methyl, ethyl, (C3-C6)alkyl, (C1-C4)alkoxy or trifluoromethyl group and with a substituent X, X being as defined above;
a 1-naphthyl or 2-naphthyl radical;
a 1-naphthyl or 2-naphthyl radical substituted in positions 5, 6, 7 and/or 8 with one or two hydroxyl groups, one or two (C1-C4)alkoxy groups or a 6,7-methylenedioxy group;
or Zxe2x80x3 is hydrogen and Z and Zxe2x80x2 represent, each independently, a non-substituted or mono-, di- or tri-substituted phenyl group;
or of one of its pharmaceutically acceptable salts and solvates, for the preparation of pharmaceutical compositions capable of increasing circulating and cellular and extracellular levels of TGF-xcex21.
According to an advantageous aspect, the object of the invention is the use of the compound of formula (I) in which Y is CH and R1 is o- or m-CF3.
According to a preferred aspect, Y is CH, R1 is o- or m-CF3 and Zxe2x80x2 and Zxe2x80x3 are hydrogen.
According to another preferred aspect, Y is CH, R1 is o- or m-CF3, Zxe2x80x2 and Zxe2x80x3 represent an oxo group and Z is 4-biphenyl.
According to a further advantageous aspect, the object of the invention is the use of the compound of formula (1) wherein Y is CH, R1 is o- or m-CF3, Zxe2x80x2 and Zxe2x80x3 are hydrogen and Z represents a phenyl radical monosubstituted with a substituent X, X being a), b), c) or one of its pharmaceutically acceptable salts and solvates.
According to another preferred aspect, the invention relates to the use of the compound of formula (I) in which Y is CH, R1 is o- or m-CF3, Zxe2x80x2 and Zxe2x80x3 are hydrogen and Z represents either a phenyl radical monosubstituted with a group Xxe2x80x2, Xxe2x80x2 being a phenyl non-substituted or substituted with 1 to 3 halogens, 1 to 3 CF3, 1 to 3 (C1-C4)alkyl, 1 to 3 (C1-C4)alkoxy, 1 to 3 cyano, 1 to 3 amino, 1 3-C4)alkylamino, 1 to 3 (C1-C4)acylamino, 1 to 3 carboxy, 1 to 3 (C1-C 4)alkoxycarbonyl, 1 to 3 aminocarbonyl, 1 to 3 mono- or di-(C1-C4)alkylaminocarbonyl, 1 to 3 amino(C1-C4)alkyl, 1 to 3 hydroxy(C1-C4)alkyl or 1 to 3 halo(C1-C4)alkyl groups; or a phenyl radical disubstituted with a substituent R2, R2 being a halogen or a hydroxy, methyl, ethyl, (C3-C6)alkyl, (C1-C4)alkoxy or trifluoromethyl group and with a substituent Xxe2x80x2, Xxe2x80x2 being as defined above, or of one of its pharmaceutically acceptable salts and solvates.
According to another preferred aspect, the invention relates to the use of the compound of formula (I) in which Y is CH, R1 is o- or m-CF3, Zxe2x80x2 and Zxe2x80x3 are hydrogen and Z is a phenyl group substituted in positions 3 and 4 with a (C1-C6)alkyl group, or of one of its pharmaceutically acceptable salts and solvates.
According to another preferred aspect, the invention relates to the use of the compound of formula (I) in which Y is CH, R1 is o- or m-CF3, Zxe2x80x3 is hydrogen and Z and Zxe2x80x2, identical, each represent a phenyl group ; a phenyl group substituted in position 2, 3 or 4 with a fluorine or chlorine atom or with a methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, trifluoromethyl, cyano, methoxy, methylthio, methylsulphonyl, ethoxy, ethylthio, ethylsulphonyl, (C1-C3)alkoxycarbonyl or di(C1-C3)alkylaminocarbonyl group; a phenyl group disubstituted in positions 2,4; 3,4; 3,5 or 2,6 with a chlorine or fluorine atom, or with a methyl, ethyl, trifluoromethyl, cyano or methoxy group; or a phenyl group trisubstituted in positions 3,4,5; 2,4,5 or 2,4,6 with a chlorine or fluorine atom, or with a methyl, ethyl, trifluoromethyl, cyano or methoxy group, or of one of its pharmaceutically acceptable salts and solvates.
According to a particularly preferred aspect, the invention relates to the use of the compound of formula (I) in which Y is CH, R1 is m-trifluoromethyl, Zxe2x80x2 and Zxe2x80x3 are hydrogen and Z represents a naphthyl, 6,7-dimethoxy-2-naphthyl or 6,7-methylenedioxy-2-naphthyl group, or of one of its pharmaceutically acceptable salts and solvates.
A particularly advantageous compound according to the present invention may be selected amongst:
1-(2-naphth-2-ylethyl)-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(6,7-dimethoxynaphth-2-yl)ethyl)-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(6,7-methylenedioxynaphth-2-yl)ethyl)-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyride ;
1-[2-(4-isobutylphenyl)propyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[(2S)-2-(4-isobutylphenyl)propyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[(2R)-2-(4-isobutylphenyl)propyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-isobutylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-tertbutylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-isobutylphenyl)-2-methylpropyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-isopropylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3xe2x80x2-chloro-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(2xe2x80x2-chloro-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4xe2x80x2-chloro-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4xe2x80x2-fluoro-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3 xe2x80x2-trifluoromethyl-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-cyclohexylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-biphenylyl)-2-ethyl]-4-(4-fluorophenyl-1,2,3,6-tetrahydropyridine;
1-[2-(4-biphenylyl)-2-methylpropyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-phenoxyphenyl)-2-ethyl]-4-(3-trifluoromethylphenyl) -1,2,3,6-tetrahydropyridine;
1-[2-(4-benzylphenyl)-2-ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-n-butylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-n-butoxyphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3,4-diethylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3-methyl-4-pentylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-methyl-3-pentylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3,4-diethylphenyl)ethyl]-4-(6-chloropyrid-2-yl)-1,2,3,6-tetrahydropyridine;
1-(2,2-diphenylethyl)-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2,2-(4,4 xe2x80x2-dichlorodiphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2,2-(3,3 xe2x80x2-bistrifluoromethyldiphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-2,2-(4,4xe2x80x2-dimethoxydiphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-fluorophenyl)-2-phenylethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-(3,3-diphenylpropyl)-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2,2-(4,4xe2x80x2-dichlorodiphenyl)ethyl]-4-(6-chloropyrid-2-yl)-1,2,3,6-tetrahydropyridine;
1-[2-(3xe2x80x2-chlorobiphenyl-4-yl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(2xe2x80x2-chlorobiphenyl-4-yl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4xe2x80x2-chlorobiphenyl-4-yl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-isobutylphenyl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-benzylphenyl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-cyclohexylphenyl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4xe2x80x2-fluorobiphenyl-4-yl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-n-butylphenyl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(biphenyl-4-yl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-t-butylphenyl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4xe2x80x2-trifluoromethylbiphenyl-4-yl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(2,3xe2x80x2-dichloro-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3-chloro-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3xe2x80x2,5xe2x80x2-dichloro-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(2xe2x80x2,4xe2x80x2-dichloro-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrabydropyridine;
1-[2-(2-chloro-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3xe2x80x2-chloro-4-biphenylyl)-2-methylpropyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(2-fluoro-4-biphenylyl)propyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-methoxy-3-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4xe2x80x2-methoxy-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4xe2x80x2-hydroxy-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4xe2x80x2-ethoxycarbonylbutoxy-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3xe2x80x2chloro-4xe2x80x2-fluoro-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(2xe2x80x2-trifluoromethyl-4-biphenylyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(biphenyl-4-yl)ethyl]-4-(2-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-cyclohexenylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3,4-diisobutylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3,4-dipropylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(4-cyclohexylphenyl)ethyl]-4-(6-chloropyrid-2-yl)-1,2,3,6-tetrahydropynidine;
1-[2-(4-isobutylphenyl)propyl]-4-(6-chloropyrid-2-yl)-1,2,3,6-tetrahydropyridine;
1-[2-(2xe2x80x2-trifluoromethylbiphenyl-4-yl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
1-[2-(3xe2x80x2-trifluoromethylbiphenyl-4-yl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine;
and their pharmaceutically acceptable salts and solvates.
1 -(2-naphth-2-ylethyl)-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine, known by its laboratory code SR 57746 and its pharmaceutically acceptable salts and solvates, especially its hydrochloride (SR 57746A), are particularly preferred compounds for the use according to the present invention.
Certain compounds of formula (I) are novel products. Thus, according to another of its aspects, the present invention relates to a compound of formula (I) selected amongst:
1-[2-(6,7-methylenedioxynaphth-2-yl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine,
1-[2-(4-cyclohexenylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine, and
1-[2-(biphenyl-4-yl)ethyl]-4-(2-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine, and their pharmaceutically acceptable salts and solvates.
The salts with pharmaceutically acceptable bases are for example those with alkali metals or alkaline earth metals, such as sodium, potassium, calcium, magnesium, and those with organic bases, such as amines, basic amino acids (lysine, arginine, histidine), trometamol, N-methylglutamine, etc.
The salts with pharmaceutically acceptable acids are for example those with mineral acids, such as hydrochloride, hydrobromide, borate, phosphate, sulphate, hydrogensulphate, hydrogenphosphate, and those with organic acids, such as citrate, benzoate, ascorbate, methylsulphate, naphthalene-2-sulphonate, picrate, fumarate, maleate, malonate, oxalate, succinate, acetate, tartrate, mesylate, tosylate, isethionate, xcex1-ketoglutarate, (xcex1-glycerophosphate, glucose-1-phosphate, etc.
The compounds of formula (I) in which Zxe2x80x2 and Zxe2x80x3 are hydrogen or a (C1-C3)alkyl group are prepared as described in WO 97/01536.
The compounds of formula (I) in which one of Zxe2x80x2 and Zxe2x80x3 is hydrogen and the other is a hydroxyl, as well as the compounds in which Zxe2x80x2 and Zxe2x80x3 together represent an oxo group, may be prepared as described in WO 93/1 1107.
The compounds of formula (I) wherein Zxe2x80x3 is hydrogen and Zxe2x80x2 and Z each represent independently a non-substituted mono-, di-, or tri-substituted phenyl group are prepared according to the following method:
(a) an aryl-1,2,3,6-tetrahydropyridine of formula (II) 
in which Y and R1 are as defined above is allowed to react with an acid of formula (III) 
in which Z and Zxe2x80x2 are as defined above, or with one of its functional derivatives,
(b) the carbonyl intermediate of formula (IV) 
is reduced, and
(c) the compound of formula (I) thus obtained is isolated and, optionally, transformed into one of its salts or solvates.
The reaction of step (a) can be conveniently carried out in an organic solvent at a temperature between xe2x88x9210xc2x0 C. and the reflux temperature of the reaction mixture; preferably the reaction is carried out at a low temperature.
The reaction solvent used is preferably a halogenated solvent such as methylene chloride, dichloroethane, 1,1,1-trichloroethane, chloroform and similar ones, or an alcohol such as methanol or ethanol, but other organic solvents which are compatible with the reagents employed, for example dioxane, tetrahydrofuran or a hydrocarbon such as hexane, may also be employed.
The reaction may conveniently be carried out in the presence of a proton acceptor, for example an alkaline carbonate or a tertiary amine. The free acid, optionally activated (with BOP for example), the anhydride, a mixed anhydride, an activated ester or an acid halide, preferably the chloride or the bromide, may be used as a suitable functional derivative of the acid of formula (III). Amongst the activated esters, the p-nitrophenyl ester is particularly preferred, but the methoxyphenyl, trityl, benzhydryl esters and similar ones are also suitable.
The reduction of step (b) may conveniently be carried out by suitable reducing agents such as aluminium hydrides or a lithium aluminium complex hydride in an inert organic solvent at a temperature between 0xc2x0 C. and the reflux temperature of the reaction mixture according to usual techniques.
xe2x80x9cInert organic solventxe2x80x9d is understood as meaning a solvent which does not interfere with the reaction. Such solvents are for example ethers, such as diethyl ether, tetrahydrofuran, dioxane or 1,2-dimethoxyethane.
The compound of formula (I) obtained is isolated according to usual techniques and optionally transformed into one of its acid addition salts or, when an acid group is present, the amphoteric character of the compound enables the separation of the salts either with acids or with bases.
The starting amines of formula (II) in which Y is CH are known compounds or may be prepared according to analogous procedures to those used for preparing the known compounds.
The starting amines of formula (II) in which Y is N may be prepared by the reaction of a suitable 2-halopyridine of formula (p) 
in which R1 is as defined above and Hal is a halogen atom, with a 1,2,3,6-tetrahydropyridine of formula (q) 
in which Po represents a protecting group such as a benzyl group for example, and Z represents a substituent which enables nucleophilic substitution of the halogen of the pyridine. Such substituents are for example trialkylstannanes, such as tributylstannane, or Grignard compounds.
The 1,2,3,6-tetrahydropyridine is then deprotected by cleaving the protecting group under suitable conditions.
The acids of formula (III) may be prepared according to the Wittig reaction by the reaction of a suitable benzophenone of formula (r) 
in which Z and Zxe2x80x2 are as defined above, with trimethylsulphoxonium iodide/BF3-Et2O and the oxidation of the intermediate aldehyde of formula (w) 
according to the method described in J. Am. Chem. Soc., 1990. 112(18):6690-6695, to obtain the corresponding acid.
According to another method, the compounds of formula (I) in which Zxe2x80x3 is hydrogen may also be prepared by the reaction of an aryl-1,2,3,6-tetrahydropyridine of formula (II) 
in which R1 and Y are as defined above, with an aldehyde of formula (w) above in the presence of a reducing agent such as sodium cyanoborohydride, according to known techniques.
The compounds of formula (I), in which R1 is m-trifluoromethyl, Y is CH, Zxe2x80x2 and Zxe2x80x3 are hydrogen and Z is a naphthyl group substituted with one or two alkoxy groups or with a methylenedioxy group, are prepared as described in EP 0 458 697.
1 -(2-naphth-2-ylethyl)-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine and its pharmaceutically acceptable salts and solvates, especially the hydrochloride, may be prepared according to EP 0 101 381.
An advantageous method provides the reaction of 2-(2-bromoethyl)naphthalene and 4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine and the isolation preferably of 1-(2-naphth-2-ylethyl)-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine hydrochloride (SR 57746A) which is then crystallised in an ethanol/water mixture by heating and cooling to 5xc2x0 C. with a cooling gradient of 10xc2x0 C. /hour and a stirring speed of 400 r.p.m., so as to obtain a mixture of the two crystalline forms in a ratio of about 66/34.
The 1 -(2-naphth-2-ylethyl)-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine hydrochloride is preferably used in a micro-particulate form, for example in an essentially amorphous form obtained by atomisation, or in a micro-crystalline form obtained by micronisation.
The effect of the compounds of formula (I) upon the increase of the levels of TGF-xcex21 was evaluated with the aid of tests upon the smooth muscle cells, as well as upon the blood levels and the diaphragms in the rat after administration of the representative compounds of the invention.
Both latent TGF-xcex21 and activated TGF-xcex21 were determined on the smooth muscle cells by incubation with hydrochloric acid.
In these tests, the representative compounds of formula (I) showed an increase in the levels of TGF-xcex21.
The anti-apoptotic activity was measured on the same cells vis-à-vis the pro-apoptotic activity of a deprivation in serum or after the addition of toxic compounds such as vincristine or growth factors such as nerve growth factor (NGF) with the aid of a specific ELISA (enzyme-linked immunosorbent assay) determination kit which detects the presence of oligonucleosomes the presence of which inside the cells is a specific marker of programmed cell death (apoptosis), according to the method described by Del Bino G. et al., (Experimental Cell Research, 193, 27, 1991 and 195, 485, 1991) or Darzynkiewicz A et al., (Cytometry, 13, 795, 1992).
In the three cases, the representative compounds of the invention, especially:
1-[2-(3,4-diethylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine (compound A);
1-[2-(biphenyl-4-yl)-2-oxoethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine (compound B);
1-[2-(biphenyl-4-yl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine (compound C);
1-[2-(6,7-methylenedioxynaphth-2-yl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine (compound D);
1-[2-(4-cyclohexenylphenyl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine (compound E)
1-[2-(biphenyl-4-yl)ethyl]-4-(2-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine (compound F); and
1-(2-naphth-2-ylethyl)-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine (SR 57746)
and their pharmaceutically acceptable salts, inhibit, as a function of time and the dose, the pro-apoptotic effect induced by the deprivation in serum or even by the addition of NGF or vincristine.
Thus, according to a further aspect, the present invention relates to the use of tetrahydropyridines of formula (I), of the advantageous or preferred compounds cited above, or of their pharmaceutically acceptable salts and solvates for the preparation of a medicament for treating diseases treatable by increasing the levels of TGF-xcex21. Such pathologies are for example diseases linked to an abnormal apoptotic activity, ocular diseases such as cataracts or glaucoma, osteoporosis, bone fractures, epidermal lesions, restenosis, conditions linked to an incorrect proliferation or migration of the smooth muscle cells, inflammations of the respiratory system, asbestosis, silicosis, lupus erythematosus, Goodpasture""s syndrome, granulomatosis, eosinophilic granulomatosis, gastric and duodenal ulcers, oesophagitis, enteritis, gastritis, septicaemia, dysfunctions of the haematopoiesis and/or lymphopoiesis, cystic fibrosis.
According to a particularly advantageous aspect, the present invention relates to the use of tetrahydropyridines of formula (I), of the advantageous or preferred compounds cited above, or of their pharmaceutically acceptable salts and solvates for the preparation of medicaments capable of inhibiting apoptosis.
It is by virtue of this anti-apoptotic activity that the compounds of the present invention may be used for the preparation of medicaments for treating cancer and its metastases, infections by antiviruses such as HIV and HTLV 1 and 2 (human immunodeficiency virus and human T lymphocyte virus) and the consequences thereof such as ATL (Adult-cell Leukaemia), leukaemia, myelopathies and arthropathies, hepatites (C, A, B, F), AIDS, immune deficiencies, cell aging, tissue degeneration phenomena, inflammation, cell proliferation, infectious diseases, graft rejection, acute or chronic rheumatoid arthritis, ulcerative colitis, thrombocytopenic purpura, autoimmune erythronoclastic anaemia, juvenile (Type I) diabetes (insulin-dependent), myelodysplasic syndrome, Huntington""s disease, prion diseases, ARDS, prostatic hypertrophy, asthma, atherosclerosis and its thrombo-embolic complications, renal diseases, glomerulonephritis, ischemic pathologies such as myocardial infarction, myocardial ischemia, coronary vasospasm, angina and cardiac failure, chronic pancreatitis, auto-immune gastritis, primary biliary cirrhosis.
According to an advantageous aspect, the present invention relates to the use of tetrahydropyridines of formula (I), of one of the advantageous or preferred compounds cited above, or of their pharmaceutically acceptable salts and solvates for the preparation of medicaments capable of treating a disease such as graft rejection or acute or chronic rheumatoid arthritis.
According to the aim of the present invention, xe2x80x9ctreatment of diseasesxe2x80x9d is understood as meaning both the treatment and the prevention of the diseases, when this is possible. Thus, for example, when graft rejection is considered, the pharmaceutical compositions may be used in the aim of prevention.
According to a further aspect, the invention relates to a method for increasing circulating and cellular and extracellular levels of TGF-xcex21.
According to another of its aspects, the present invention relates to a method for inhibiting apoptosis, which comprises the administration to a mammal in need thereof of an effective dose of a compound of formula (I), of one of the advantageous or preferred compounds cited above, or of one of their pharmaceutically acceptable salts and solvates, advantageously SR 57746, or one of its pharmaceutically acceptable salts and solvates.
According to a preferred aspect, SR 57746 and its pharmaceutically acceptable salts and solvates are administered in a micro-particulate form, preferably in a micro-particulate form of the hydrochloride.
The compounds of formula (I), one of the advantageous or preferred compounds cited above or their pharmaceutically acceptable salts and solvates are preferably administered orally.
The amount of active principle to be administered depends upon the degree of advancement of the disease as well as the age and weight of the patient. However, the unit doses generally comprise from 0.25 to 700 mg, advantageously from 0.5 to 300 mg, preferably from 1 to 150 mg, for example between 2 and 50 mg of active principle. These unit doses are normally administered once or more times a day, preferably once to three times per day, the overall dose in man being variable between 0.5 and 1,400 mg per day, for example from 1 to 900 mg per day, advantageously from 2 to 500 mg per day, more conveniently from 2 to 200 mg per day. When the active principle administered is for example SR 57746, the unit dose generally comprises from 0.5 to mg, advantageously from 1 to 5, preferably from 1 to 3 mg, for example 1-1.5-2-2.5-3 mg of active principle. These unit doses are normally administered once or more times per day, preferably once to three times per day, the overall dose in man being variable between 0.5 and 50 mg per day, for example from 1 to mg per day, advantageously from 2 to mg per day.
The doses and amounts above refer to the compounds of formula (I) or to one of the advantageous or preferred compounds cited above, in a non-salified form.
In the pharmaceutical compositions of the present invention for oral administration, the active principle may be administered as unit forms for administration, in a mixture with classical pharmaceutical carriers, to mammals, to animals and to human beings for the treatment of the above-mentioned diseases. The suitable unit forms of administration comprise for example tablets, which are optionally scored, gelatine capsules, powders, granules and oral solutions or suspensions.
When a solid composition is prepared in the form of tablets, the main active ingredient is mixed with a pharmaceutical vehicle such as gelatine, starch, lactose, magnesium stearate, talc, gum arabic or analogues thereof. The tablets may be coated with sucrose or other suitable materials or even they may be treated such that they have a sustained or delayed activity and that they continuously release a predetermined amount of the active principle.
A gelatine capsule preparation is obtained by mixing the active ingredient with a diluant and in pouring the mixture obtained into soft or hard gelatine capsules.
A preparation in the form of a syrup or elixir may contain the active ingredient together with a sweetener, preferably an acalorific sweetener, methylparaben and propylparaben as antiseptics, as well as a flavouring agent and a suitable colouring agent.
Powders or granules which may be dispersed in water can contain the active ingredient in a mixture with dispersing agents or wetting agents, or suspension agents, such as polyvinylpyrrolidone, as well as with sweeteners or flavour correctors.
The active principle may also be formulated in the form of microcapsules, optionally with one or more carriers or additives.
In the pharmaceutical compositions according to the present invention, the active principle may also be in the form of an inclusion complex in cyclodextrins, their ethers or their esters. The PREPARATIONS and EXAMPLES below illustrate the invention better.
40,000 smooth muscle cells isolated from the human aorta (supplier: CLONETICS) are placed, in a mm dish, in a medium containing 2 ml of DMEM (Dulbecco Modified Eagle Medium containing 4.5 g/l of glucose, 3.7 g/l of NaHCO3 and not containing any L-glutamine or Na-pyruvate). 20% v/v of foetal calf serum which was desupplemented for 30 min at 80xc2x0 C. , 4 mM L-glutamine, 50 U/ml of penicillin and 50 xcexcg/ml of streptomycin are added. The cells are left in this medium for a growth period of three days before submitting them to tests according to the examples given further on.
Dishes containing cells are prepared as described in Preparation 1. Apoptosis is induced by replacing the medium described in Preparation 1 with the same medium containing only 0.2% of foetal calf serum. The effect of the compounds of the invention upon the levels of latent and activated TGF-xcex21 is measured in the extracellular media after 24 hours of contact with the cells, in comparison with controls (0.2% of foetal calf serum and 20% of foetal calf serum). The activated TGF-xcex21 is determined directly in the supernatants of the culture, but the latent TGF-xcex21 is determined after activation. For the activation, 0.5 ml of the supernatant of the culture are incubated in the presence of 0.1 ml of 1M HCl for 10 min. at room temperature. The mixture is then neutralised with 0.1 ml of 0.5 M Hepes buffer which contains 1.2 M NaOH. Determinations of TGF-xcex21 are carried out with the aid of a specific ELISA test.
5 Sprague Dawley rats (Iffa Credo, France) of about 280 g were treated every day for 15. three days with the compound to be tested, which was administered orally, 24 hours after the last forced feeding, the rats are anaesthetised. Blood is taken from the abdominal aorta on EDTA, the samples are centrifuged and the supernatants (plasma rich in platelets) are frozen. The diaphragms are also taken, rinsed several times in cold PBS (Phosphate buffered saline) and are centrifuged. After a further ultracentrifugation the plugs are taken up into PBS and frozen. Determinations of latent and activated TGF-xcex21 in the plasma and the ground diaphragms are made by using the technique described in PREPARATION 2. The increase in the circulating latent TGF-xcex21 levels are recorded in the rats treated with the compounds of the invention in comparison with control rats. The increase in the activated TGF-xcex21 levels in the diaphragms of the rats treated with the compounds of the invention are also recorded.
Dishes containing the cells are prepared as in PREPARATION 1. Apoptosis is induced by three different methods:
a) by replacing the medium of PREPARATION 1 with the same medium containing only 0.2% of foetal calf serum;
b) by adding increasing doses of NGF (0.01 ng/ml to 100 ng/ml) to the medium described in PREPARATION 1;
c) by adding increasing doses of Vincristin (0.1 pg/ml to 10 ng/ml) to the medium described in PREPARATION 1.
By an ELISA determination test of the mono- and oligo-nucleosomes associated with cytoplasmic histones after washing and cellular lysis, the effects of the compounds of the invention upon the apoptosis are measured after 24 hours of contact with the cells in comparison with the levels of apoptosis obtained in the absence of products (maximum apoptosis level) or in the presence of 20% foetal calf serum (minimum apoptosis level).